Device and method for fragmenting polymers and particles

Abstract

The invention features devices for fragmenting polymers or particles in a sample using a centrifuge. In its simplest embodiment, the device includes a body and a valve. The valve, to one side of which a sample is applied, is designed to remain closed until a specified centrifugal force is reached or exceeded. Once open, the valve allows the passage of the sample through a channel in the body. The channel includes one to four fragmenting regions, and the polymers or particles are fragmented as they traverse the fragmenting regions under an applied centrifugal force. The invention further features methods of fragmenting polymers using a device of the invention.

What is claimed is: 
     
         1 . A device for fragmenting polymers or particles in a liquid sample, the device comprising a body adapted for use in a centrifuge;
 a valve; and   a channel formed in the body and having a first end and a second end, wherein one to four fragmenting regions are disposed between the first and second ends,   wherein the valve opens thereby allowing the sample to pass through the valve and channel, and the polymers or particles are fragmented as they traverse the one to four fragmenting regions under a centrifugal force.   
     
     
         2 . The device of  claim 1 , wherein (i) the body further comprises a sample compartment, and the sample is substantially retained in the sample compartment until the valve opens; (ii) the body is adapted to be supported in a centrifuge tube, (iii) placement of the device in the centrifuge tube defines a sample compartment, and the sample is substantially retained in the sample compartment until the valve opens; (iv) the body further comprises a compartment to receive fragmented molecules or particles passing through the channel; or (v) the body is shaped for placement in a centrifuge in the absence of a centrifuge tube. 
     
     
         3 - 4 . (canceled) 
     
     
         5 . The device of  claim 1 , wherein the valve: (i) opens at or above the centrifugal force; (ii) is a frangible barrier or a spring-loaded valve; (iii) is a plug that is released from the body at or above the centrifugal force; or (iv) is electronically controlled, temperature controlled, or time controlled. 
     
     
         6 - 8 . (canceled) 
     
     
         9 . The device of  claim 1 , wherein the polymers are polynucleotides or polypeptides, or the particles are cells, cellular organelles, lipid particles, or micelles that are fragmented as they traverse the one to four fragmenting regions. 
     
     
         10 . The device of  claim 9 , wherein the polynucleotides are fragmented on average into pieces ranging from 100 by to 40,000 bp. 
     
     
         11 . (canceled) 
     
     
         12 . The device of  claim 1 , wherein each fragmenting region is a narrowed region of the channel disposed at or adjacent to the second end, and the narrowed region optionally has a diameter of between 1 and 1,000 μm. 
     
     
         13 . (canceled) 
     
     
         14 . The device of  claim 1 , wherein each fragmenting region comprises a plurality of narrowed regions of the channel disposed at or adjacent to the second end. 
     
     
         15 . (canceled) 
     
     
         16 . The device of  claim 1 , further comprising a filter disposed to contact the fragmented sample after exiting the second end, wherein optionally (i) the filter is a size-exclusion filter, (ii) the filter comprises an affinity reagent for binding a component of the fragmented sample, (iii) the device further comprises at least an additional filter wherein the filters are disposed to collect fragments in a specified size range, or (iv) the filter is within the body. 
     
     
         17 - 20 . (canceled) 
     
     
         21 . The device of  claim 1 , further comprising an agent selected from the group consisting of enzymes, solvents, detergents, and biological agents to disrupt particles, wherein optionally (i) the agent is stored in liquid or dry form in the device, (ii) the device further comprises a disrupting compartment in which the agent is stored, or (iii) the device further comprises a filter to separate debris from particle disruption. 
     
     
         22 - 23 . (canceled) 
     
     
         24 . The device of  claim 1 , further comprising an acoustic transducer for sonication of the sample, electrodes for delivery an electric field to the sample, a heater or cooler, or a homogenizer having rotory cutting or chopping action. 
     
     
         25 - 26 . (canceled) 
     
     
         27 . A device comprising a device of  claim 1  placed in a centrifuge tube. 
     
     
         28 . A kit comprising a plurality of devices of  claim 1  and a holder adapted for supporting the plurality of devices in a centrifuge. 
     
     
         29 . A method for fragmenting polymers or particles in a liquid sample, said method comprising the steps of:
 a) providing a device comprising:
 (i) a body adapted for use in a centrifuge; 
 (ii) a valve; and 
 (iii) a channel formed in the body and having a first end and a second end, wherein one to four fragmenting regions are disposed between the first and second ends, 
   wherein either the body further comprises a sample compartment, or placement of the device in the centrifuge tube defines a sample compartment   wherein the sample is substantially retained in the sample compartment until the valve opens;   b) placing the sample in the sample compartment;   c) centrifuging the device at a centrifugal force; and   d) allowing the valve to open or opening the valve, wherein the sample passes through the valve and channel, and the polymers or particles are fragmented as they traverse the one to four fragmenting regions.   
     
     
         30 . The method of  claim 29 , wherein the valve: (i) opens at or above the centrifugal force, (ii) is a frangible barrier or a spring-loaded valve, (iii) is released from the body at or above the centrifugal force, or (iv) is electronically controlled, temperature controlled, or time controlled. 
     
     
         31 - 33 . (canceled) 
     
     
         34 . The method of  claim 29 , wherein the polymers are polynucleotides or polypeptides or the particles are cells that are fragmented as they traverse the one to four fragmenting regions. 
     
     
         35 . The method of  claim 34 , wherein the polynucleotides are fragmented on average into pieces ranging from 100 bp to 40,000 bp. 
     
     
         36 . (canceled) 
     
     
         37 . The method of  claim 29 , wherein each fragmenting region is a narrowed region of the channel disposed at or adjacent to the second end, and optionally the narrowed region has a diameter of between 1 and 1,000 μm. 
     
     
         38 . (canceled) 
     
     
         39 . The method of  claim 29 , wherein each fragmenting region comprises a plurality of narrowed regions of the channel disposed at or adjacent to the second end. 
     
     
         40 . The method of  claim 29 , wherein the centrifugal force is between and 1,000 and 200,000 G. 
     
     
         41 . The method of  claim 29 , wherein steps b)-d) are repeated on the same sample. 
     
     
         42 . The method of  claim 29 , wherein the device further comprises a filter disposed to contact the fragmented sample after exiting the second end, and optionally (i) the filter concentrates the fragmented sample, (ii) the filter comprises an affinity reagent for binding a component of the fragmented sample, or (iii) the device further comprises at least an additional filter wherein the filters are disposed to collect fragments in a specified size range. 
     
     
         43 - 45 . (canceled) 
     
     
         46 . The method of  claim 29 , further comprising contacting the sample with an agent selected from the group consisting of enzymes, solvents, detergents, and biological agents to disrupt particles. 
     
     
         47 . The method of  claim 29 , further comprising disrupting particles in the sample with sonication, electric field, heat, cold, or a homogenizer having rotory cutting or chopping action, and optionally the device further comprises a filter to separate debris from particle disruption from the sample to be fragmented. 
     
     
         48 . (canceled)